● Efficient and rapid: rapid ligation within 5min, and the positive rate was nearly 100%.

● Broad sensitivity: Suitable for efficient ligation of fragments as low as 0.025pmol and fragments up to 3 kb.

● It is easy to operate: With the use of the new RapiLigation Mix, the ligation reaction can be performed simply by adding the vector and fragment.

The pLB-T Fast rapid cloning kit provides efficient cloning of a wide range of PCR products and any DNA fragment with a sticky end. The kit is effective for both phosphorylate and non-phosphorylated DNA fragments. More than 95% of the positive recombinant clones were obtained in only 5min by ligation of the positive selection vector and the insert. The kit is equipped with a new type of RapiLigation Mix as a high-efficiency reagent for T4 DNA ligase reaction, which contains a linking enhancer and an enzyme stabilizer, thus greatly shortening the linking time and improving the linking and cloning efficiency of PCR products. According to different needs, DH5α competent cells and the control supercoiled plasmid were equipped for convenient transformation.

PLB-T Vector map

PLB-T Vector polyclonal site

● The 3' end of the PCR fragment to be used for ligation should have an A-terminal. In the case of a flat-ended fragment without an A-terminal that is amplified using a high-fidelity polymerase such as pfu, the pLB zero-background rapid cloning kit or the pGM-T flat-ended ligation kit can be selected for the ligation reaction.

● It is very necessary to use Control Insert DNA as the control during the transformation, and the cause can be determined when there is any problem in the experiment.

● It is recommended that a portion of the linker be left behind for rapid resolution of the problem and reduction of unnecessary repeat experiments.

● The coating amount can be adjusted according to the specific experiment. If the total amount of transformed DNA is greater, a smaller amount of the transformed product may be coated on the plate; Conversely, if the total amount of DNA converted is small, 200 to 300 L of the conversion product may be coated on the plate. If fewer clones were expected, part of the medium could be removed by centrifugation (4000 rpm, 2 min) and a suitable amount of medium was left to suspend the cells and applied to a plate (the remaining applied liquid could be stored at 4 °C; if the number of transformed colonies on the next day was too small, the remaining liquid could be applied to a new plate).